Nuclear matrix proteins, polynucleotide sequences encoding them, and their uses

Abstract

Nuclear matrix proteins (NMP) which are characterized by a defined expression in tissue, particularly prostate tissue, are provided. These NMPs are useful markers in diagnosing and monitoring the stage of malignancy of a cell and treating cell proliferative disorders associated with the NMP. Also provided are substantially purified polypeptides and nucleotide sequences encoding the NMPs of the invention.

What is claimed is:  
     
         1 . A method of determining the presence or absence of a metastatic cell, comprising determining the presence or absence of a nuclear matrix protein that is present in a metastatic cell but absent in a non-metastatic cell.  
     
     
         2 . The method as claimed in  claim 1 , wherein said determination step is performed on a sample of prostate cells.  
     
     
         3 . The method as claimed in  claim 2 , wherein the nuclear matrix protein is AM-1 having a molecular weight of about 40 kD and a pI of about 6.73, which is present in a metastatic prostate cell but absent in a non-metastatic prostate cell.  
     
     
         4 . The method as claimed in  claim 3 , wherein said determination step is performed with an antibody directed against AM-1 or an immunogenic fragment thereof.  
     
     
         5 . A purified nuclear matrix protein which is present in cancerous prostate cells but absent in normal prostate cells selected from the group consisting of: 
 (a) D-1 having a molecular weight of about 63 kD and a pI of about 8.55,    (b) D-2 having a molecular weight of about 40 kD and a pI of about 5.91, and    (c) D-3 having a molecular weight of about 33 kD and a pI of about 6.97;    or an immunogenic fragment of one of said proteins (a)-(c).    
     
     
         6 . The protein of  claim 5 , wherein the protein is D-1 having a molecular weight of about 63 kD and a pI of about 8.55.  
     
     
         7 . The protein of  claim 6 , which includes at least one sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.  
     
     
         8 . The protein of  claim 5 , wherein the protein is D-2 having a molecular weight of about 40 kD and a pI of about 5.91.  
     
     
         9 . The protein of  claim 8 , which includes at least one sequence selected from the group consisting of SEQ ID NO: 4 and SEQ ID NO: 5.  
     
     
         10 . The protein of  claim 5 , wherein the protein is D-3 having a molecular weight of about 33 kD and a pI of about 6.97.  
     
     
         11 . The protein of  claim 10 , which includes SEQ ID NO: 6.  
     
     
         12 . A method of producing an antibody that differentiates between cancerous and normal prostate cells, comprising 
 selecting a protein or fragment of  claim 5  as an antigen,    raising an antibody against said antigen, and    recovering said antibody.    
     
     
         13 . The method as claimed in  claim 12 , wherein the antibody is a monoclonal antibody.  
     
     
         14 . A purified nuclear matrix protein that is present in normal prostate cells but absent in cancerous prostate cells, and that is selected from the group consisting of: 
 (a) NDP-1 having a molecular weight of about 95 kD and a pI of about 6.74,    (b) NDP-2 having a molecular weight of about 57 kD and a pI of about 8.33,    (c) NDP-3 having a molecular weight of about 57 kD and a pI of about 8.0,    (d) NDP-4 having a molecular weight of about 47 kD and a pI of about 5.26,    (e) NDP-5 having a molecular weight of about 47 kD and a pI of about 5.80,    (f) NDP-6 having a molecular weight of about 41 kD and a pI of about 6.83,    (g) NDP-7 having a molecular weight of about 37.2 kD and a pI of about 7.05,    (h) NDP-8 having a molecular weight of about 36.9 kD and a pI of about 7.35,    (i) NDP-9 having a molecular weight of about 35 kD and a pI of about 6.25, and    (j) NDP-10 having a molecular weight of about 32.5 kD and a pI of about 5.46;    or an immunogenic fragment of one of said proteins (a)-(j).    
     
     
         15 . The protein of  claim 14 , which is NDP-1 having a molecular weight of about 95 kD and a pI of about 6.74.  
     
     
         16 . The protein of  claim 14 , which is NDP-2 having a molecular weight of about 57 kD and a pI of about 8.33.  
     
     
         17 . The protein of  claim 14 , which is NDP-3 having a molecular weight of about 57 kD and a pI of about 8.0.  
     
     
         18 . The protein of  claim 14 , which is NDP-4 having a molecular weight of about 47 kD and a pI of about 5.26.  
     
     
         19 . The protein of  claim 14 , which is NDP-5 having a molecular weight of about 47 kD and a pI of about 5.80.  
     
     
         20 . The protein of  claim 14 , which is NDP-6 having a molecular weight of about 41 kD and a pI of about 6.83.  
     
     
         21 . The protein of  claim 14 , which is NDP-7 having a molecular weight of about 37.2 kD and a pI of about 7.05.  
     
     
         22 . The protein of  claim 14 , which is NDP-8 having a molecular weight of about 36.9 kD and a pI of about 7.35.  
     
     
         23 . The protein of  claim 14 , which is NDP-9 having a molecular weight of about 35 kD and a pI of about 6.25.  
     
     
         24 . The protein of  claim 14 , which is NDP-10 having a molecular weight of about 32.5 kD and a pI of about 5.46.  
     
     
         25 . A method of producing an antibody that differentiates between cancerous and normal prostate cells, comprising 
 selecting a protein or fragment of  claim 15  as an antigen,    raising an antibody against said antigen, and    recovering said antibody.    
     
     
         26 . The method as claimed in  claim 25 , wherein the antibody is a monoclonal antibody.  
     
     
         27 . A purified nuclear matrix protein which is present in metastatic prostate cells but absent in non-metastatic prostate cells selected from the group consisting of: 
 (a) AM-1 having a molecular weight of about 40 kD and a pI of about 6.73, and    (b) AM-2 having a molecular weight of about 36 kD and a pI of about 8.33;    or an immunogenic fragment of one of said proteins (a) or (b).    
     
     
         28 . The protein of  claim 27 , which is AM-1 having a molecular weight of about 40 kD and a pI of about 6.73.  
     
     
         29 . The protein of  claim 28 , which includes SEQ ID NO: 7.  
     
     
         30 . The protein of  claim 27 , which is AM-2 having a molecular weight of about 36 kD and a pI of about 8.33.  
     
     
         31 . The protein of  claim 30 , which includes SEQ ID NO: 8.  
     
     
         32 . A method of producing an antibody that differentiates between metastatic and non-metastatic prostate cells, comprising 
 selecting a protein or fragment of  claim 29  as an antigen,    raising an antibody against said antigen, and    recovering said antibody.    
     
     
         33 . The method as claimed in  claim 32 , wherein the antibody is a monoclonal antibody.  
     
     
         34 . A purified nuclear matrix protein which is absent in metastatic prostate cells but present in non-metastatic prostate cells selected from the group consisting of: 
 (a) G-1 having a molecular weight of about 55 kD and a pI of about 6.48, and    (b) G-2 having a molecular weight of about 52 kD and a pI of about 6.93;    or an immunogenic fragment of one of said proteins (a) or (b).    
     
     
         35 . The protein of  claim 34 , which is G-1 having a molecular weight of about 55 kD and a pI of about 6.48.  
     
     
         36 . The protein of  claim 34 , which is G-2 having a molecular weight of about 52 kD and a pI of about 6.93.  
     
     
         37 . A method of producing an antibody that differentiates between metastatic and non-metastatic prostate cells, comprising 
 selecting a protein or fragment of  claim 34  as an antigen,    raising an antibody against said antigen, and    recovering said antibody.    
     
     
         38 . The method as claimed in  claim 37 , wherein the antibody is a monoclonal antibody.  
     
     
         39 . An antibody directed against a purified nuclear matrix protein or an immunogenic fragment thereof, said protein being selected from the group consisting of: 
 (i) a protein which is present in cancerous prostate cells but absent in normal prostate cells selected from the group consisting of: 
 (a) D-1 having a molecular weight of about 63 kD and a pI of about 8.55,  
 (b) D-2 having a molecular weight of about 40 kD and a pI of about 5.91, and  
 (c) D-3 having a molecular weight of about 33 kD and a pI of about 6.97;  
   (ii) a protein which is present in normal prostate cells but absent in cancerous prostate cells selected from the group consisting of: 
 (a) NDP-1 having a molecular weight of about 95 kD and a pI of about 6.74,  
 (b) NDP-2 having a molecular weight of about 57 kD and a pI of about 8.33,  
 (c) NDP-3 having a molecular weight of about 57 kD and a pI of about 8.0,  
 (d) NDP-4 having a molecular weight of about 47 kD and a pI of about 5.26,  
 (e) NDP-5 having a molecular weight of about 47 kD and a pI of about 5.80,  
 (f) NDP-6 having a molecular weight of about 41 kD and a pI of about 6.83,  
 (g) NDP-7 having a molecular weight of about 37.2 kD and a pI of about 7.05,  
 (h) NDP-8 having a molecular weight of about 36.9 kD and a pI of about 7.35,  
 (i) NDP-9 having a molecular weight of about 35 kD and a pI of about 6.25, and  
 (j) NDP-10 having a molecular weight of about 32.5 kD and a pI of about 5.46;  
   (iii) a protein which is present in metastatic prostate cells but absent in non-metastatic prostate cells selected from the group consisting of: 
 (a) AM-1 having a molecular weight of about 40 kD and a pI of about 6.73, and  
 (b) AM-2 having a molecular weight of about 36 kD and a pI of about 8.33; and  
   (iv) a protein which is absent in metastatic prostate cells but present in non-metastatic prostate cells selected from the group consisting of: 
 (a) G-1 having a molecular weight of about 55 kD and a pI of about 6.48, and  
 (b) G-2 having a molecular weight of about 52 kD and a pI of about 6.93.  
   
     
     
         40 . A method of determining the presence or absence of a prostate cancer cell, comprising determining the presence or absence of a protein of  claim 5 .  
     
     
         41 . The method as claimed in  claim 40 , wherein said determination step is performed with an antibody directed against a protein or fragment of  claim 5.